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Zaocys dhumnades capillary electrophoresis DNA fingerprint spectrum and identification method

A DNA fingerprinting and identification method technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problem of difficult to meet the accurate, rapid and objective requirements of identification, unsatisfactory effect of physical and chemical character identification, and long identification time. and other problems, to achieve the effect of easy automatic analysis, automatic analysis, good repeatability and stability

Inactive Publication Date: 2018-02-16
BEIHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because of its high cost of breeding, high price, and huge demand, there are often counterfeit products in the commercially available medicinal materials
[0003] There are many current quality evaluation methods for black-headed snakes. The traditional identification is based on the difference in traits and physical and chemical methods, but it is highly subjective, and the identification time is long and the cost is high.
Moreover, the plasticity of the traits of the black snake in the process of drying and making decoction pieces is very large, resulting in unsatisfactory identification of physical and chemical properties, and it is difficult to meet the accurate, fast and objective requirements for the identification of the black snake

Method used

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  • Zaocys dhumnades capillary electrophoresis DNA fingerprint spectrum and identification method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Establishment of capillary electrophoresis DNA fingerprint of black snake

[0028] 1. DNA extraction

[0029] Take 0.5g of this product, put it in a mortar, grind it thoroughly to make it into powder, take 0.1g and put it in a 1.5ml centrifuge tube, add 275μl of digestive solution, keep it warm in a water bath at 55°C for 1 hour, add 250μl of lysis buffer, mix well, Add it to the DNA purification column, centrifuge (10000 rpm / 1 minute) for 3 minutes; discard the filtrate, add 800 μl of eluent, and centrifuge (10000 rpm / 1 minute) for 1 minute; discard the filtrate, and use the above elution The solution was repeatedly eluted 3 times, and centrifuged (10000 rpm / 1 min) for 1 minute each time; the filtrate was discarded, and centrifuged for 2 minutes, and the DNA purification column was transferred to another centrifuge tube, and 100 μl of sterile double distilled water was added. After standing at room temperature for 2 minutes, centrifuge (10,000 rpm / 1 minute) for 2 minu...

Embodiment 2

[0040] Applying the black-snake DNA fingerprint identification method of the present invention to the identification of a certain black-snake sample

[0041] Assuming that the existing batch of black snake samples cannot determine whether it is a real black snake, therefore, it is identified according to the following method. After the establishment of the DNA standard fingerprint of the black snake snake, when a sample is to be identified, the mitochondrial DNA is first extracted by the salting-out method; the extracted DNA is used as a template, and PCR amplification is carried out with SSR primers; the amplified product is analyzed. Capillary electrophoresis analysis and record the electrophoresis pattern, that is, the DNA fingerprint of the batch of samples to be tested; use the similarity analysis software to compare the similarity between the DNA fingerprint of the sample to be tested and the DNA standard fingerprint of the black snake snake, and according to the similari...

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Abstract

The invention relates to a traditional Chinese medicine zaocys dhumnades DNA identification technology, in particular to an SSR fluorescence labeled zaocys dhumnades capillary electrophoresis DNA fingerprint spectrum and identification method. The method has the characteristics of being high in resolution, convenient to operate, quick, capable of obtaining accurate results and the like and can beused for identifying zaocys dhumnades samples.

Description

technical field [0001] The invention relates to identification technology of traditional Chinese medicinal materials, in particular to SSR fluorescence-marked Snake DNA capillary electrophoresis fingerprint and identification method. Background technique [0002] Black-headed snake is a traditional precious medicinal material in my country. The "Chinese Pharmacopoeia" (2015 edition) stipulates that its authentic product is the dried body of the snake snake Zaocysdhumnades (Cantor). Most of them are caught in summer and autumn, the abdomen is cut open or the skin is peeled to keep the head and tail, the viscera are removed, the disk is made into a disk, and dried. It has the effects of expelling wind, dredging collaterals, and relieving spasm. For rheumatic obstinate numbness, numbness and spasm, apoplexy, hemiplegia, tetanus, etc. Because of its high cost of cultivation, high price, and huge demand, there are often counterfeit products in the commercially available medicin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/686C12Q2563/107C12Q2565/125C12Q2525/151
Inventor 苑广信李梓僮张丽华王冰梅付桂莲李盈诺孙云朋王艳双周亭亭王雪松张悦杨雪
Owner BEIHUA UNIV
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